Cholera vaccine candidate 638: intranasal immunogenicity and expression of a foreign antigen from the pulmonary pathogen Coccidioides immitis.
Cholera vaccine candidate 638: intranasal immunogenicity and expression of a foreign antigen from the pulmonary pathogen Coccidioides immitis.
复制标题
霍乱候选疫苗 638：鼻内免疫原性和来自肺部病原体粗球孢子菌的外源抗原的表达。
DOI：
10.1016/s0264-410x(03)00515-2
复制
发表时间：
2003
期刊：
影响因子：
5.5
通讯作者：
Benitez,JorgeA
中科院分区：
文献类型：
--
作者：
Silva,AnisiaJ;Mohan,Archana;Benitez,JorgeA
霍乱弧菌638株是一种遗传减毒的候选霍乱活疫苗，其中编码霍乱毒素的CTXΦ原噬菌体已被删除，编码胞外锌依赖性金属蛋白酶的hapA被插入灭活。当通过鼻内途径接种至成年瑞士小鼠时，菌株638具有高度免疫原性，这通过诱导强血清杀弧菌抗体应答来判断。在上述模型中，菌株638与其同基因hapA+前体（菌株81）的并行比较表明，hapA的灭活不影响免疫原性。粗球孢子菌球相关抗原2/富含脯氨酸抗原（Ag 2/PRA）已被证明在一定程度上依赖于抗原呈递和C.肠炎关节孢子。在这项工作中，我们证明了使用活的遗传减毒霍乱弧菌菌株提供Ag 2/PRA。利用Tac强启动子，将Ag 2/PRA与大肠杆菌不耐热毒素B亚基前导肽融合表达于638中。重组Ag 2/PRA在大肠杆菌中高效表达、加工并分泌至周质间隙。用表达Ag 2/PRA的菌株638鼻内免疫成年小鼠诱导对载体菌株的血清杀弧菌抗体应答和对Ag 2/PRA的血清总IgG应答。表达PRA的菌株638在免疫后20 h内可从气管和肺中回收，但在72 h后被有效清除。
Vibrio cholerae strain 638 is a live genetically attenuated candidate cholera vaccine in which the CTXΦ prophage encoding cholera toxin has been deleted and hapA, encoding an extracellular Zn-dependent metalloprotease, was insertionally inactivated. Strain 638 was highly immunogenic when inoculated to adult Swiss mice by the intranasal route as judged by the induction of a strong serum vibriocidal antibody response. A side-by-side comparison of strain 638 with its isogenic hapA+precursor (strain 81) in the above model indicated that inactivation of hapA does not affect immunogenicity. The spherule-associated antigen 2/proline-rich antigen (Ag2/PRA) of Coccidioides immitis has been shown to protect mice against coccidioidomycosis to an extent dependent on the modes of antigen presentation and challenge with C. immitis arthrospores. In this work, we demonstrate the use of a live genetically attenuated V. cholerae strain to deliver Ag2/PRA. Ag2/PRA was expressed in 638 as a fusion protein with the Escherichia coli heat labile toxin B subunit leader peptide using the strong Tac promoter. The recombinant Ag2/PRA was efficiently expressed, processed and secreted to the periplasmic space. Intranasal immunizations of adult mice with strain 638 expressing Ag2/PRA induced serum vibriocidal antibody response to the vector strain and serum total IgG response to Ag2/PRA. Strain 638 expressing PRA could be recovered from trachea and lung up to 20h after immunization but was effectively cleared 72h post-inoculation.